CN104083759A - Microneedle array vaccine adjuvant transmission system built by using lipid modifying carrier - Google Patents
Microneedle array vaccine adjuvant transmission system built by using lipid modifying carrier Download PDFInfo
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Abstract
The invention discloses a microneedle array which contains lipid modifying carrier and is used for a vaccine adjuvant transmission system. The microneedle array comprises a substrate and a plurality of microneedles fixed on the substrate, wherein the substrate is composed of saccharides, povidone, cellulose, or starch auxiliary materials; each microneedle comprises lipid A modifying carrier, the auxiliary materials (excipients) and vaccine ingredients; the lipid A modifying carrier can be lipidosome, lipid, micro-capsule or nanoparticle, and the like; the vaccine ingredients mainly refer to causative agent antigen proteins. Compared with the existing vaccine preparations, the lipid A-modified lipidosome microneedle array vaccine adjuvant transmission system can contain different vaccine ingredients to form vaccines aiming to different pathogens and therefore has a wide application scope. Biodegradable materials are selected and therefore the safety is high. The microneedle array vaccine adjuvant transmission system is a solid preparation which is high in stability and convenient to inoculate. Inoculation can be completed by a user self through oral mucosa, the vaccine can be prevented from running away along with saliva, and a body can be induced to establish mucosal immunity.
Description
Technical field
The vaccine adjuvant field that the present invention relates to prophylaxis against infection diseases, relates in particular to new vaccine adjuvant transmission system, is in the nature and adopts function carrier to build microneedle array, as vaccine adjuvant transmission system.
Background technology
Vaccine is the immunogenic substances that can produce antibody, is the preparation for keeping off infection.Through years development, vaccine point mainly comprises as Types Below from originating at present: 1) inactivated vaccine is selected antibacterial that immunogenicity is good, virus, rickettsia, the inferior body of spiral shell etc., through artificial culture, then is killed and makes by physics or chemical method.2) artificial site directed mutagenesis method for attenuated live vaccine, or filter out virulence attenuation of or substantially nontoxic viable microbial is made live vaccine from nature.As bacillus calmette-guerin vaccine (BCG, tuberculosis), Measles Vaccine, poliomyelitis vaccine (poliomyelitis) etc.3) RNA/DNA vaccine RNA/DNA vaccine is that the expressing gene of proteantigen is cloned on expression vector, is injected in vivo, make its antigen express in vivo rear excitating organism and produce immunoreation, and its preparation process complexity, safety is lower.4) toxoid (extracellular toxin) extracellular toxin loses toxicity after formaldehyde treated, still retains immunogenicity, is toxoid.Wherein add appropriate aluminum phosphate and aluminium hydroxide adsorption refining toxoid.5) subunit vaccine (component vaccine) is removed the composition (especially hereditary material) being even harmful to without immunization in pathogen, retains the vaccine that its effective immunogenic components is made.
How vaccine is by traditional injection inoculation at present, and systemic immunity induction efficiency is high; But side effect is many, production cost is high, needs professional inoculation personnel, has reduced vaccine prevention efficiency and cannot induce body to produce mucosal immune response.
Solvable micropin vaccine can efficiently transmit Ag, and painless, action time is longer, easy to use, gets most of the attention recently; But micropin vaccine mostly is skin patch at present, is also difficult to bring out mucosal immunity; Lack the transmission of immunocyte specificity; A little less than adjuvant function.
Comparatively speaking, mucosal vaccine has some clear superiorities: 1) tract contains a large amount of mucosa associated lymphoid tissues (MALT), mucosal vaccination can produce systemic immunity and reply, and also can produce widely (inoculation position and far-end) mucosal immune response; Because mucosa is pathogen invasion main path, therefore mucosal immunity can form two defence lines, road to pathogen; 2) have no mechanical damage, easy to use, eliminate the equipment such as injection and pollution thereof, save professional's training cost, be easy to promote; 3) production cost is lower.Therefore the various mucosal vaccines of people's develop actively.
Oral vaccine is easy to use, but needs larger dose, both increased cost and also easily caused antigen tolerance, and gastrointestinal tract severe rugged environment more becomes the obstacle that is difficult to go beyond.There is cystic fibrosis danger, Pfizer in pulmonary administration
(inhaled Insulin) just therefore withdraws from market [29].Nasal membrane is rich in MALT and is but close to central nervous system (CNS), and inoculation risk is higher, for example,
cause part inoculator facial paralysis (Bell's palsy), withdraw from subsequently market.
Oral environment relaxes, and is beneficial to and keeps Ag activity; Sublingual inoculation can produce the preventive effect that is equal to intranasal inoculation, but can not produce CNS toxicity.But, oral mucosa coating stratified squamous epithelium, and MALT is relatively far away apart from inoculation surface; And saliva and swallow and also cause Ag to run off in a large number needs anesthetized animal when inoculation, completely infeasible concerning people.
In addition, development mucosal vaccine also exists some general obstacles: after 1) subunit vaccine Ag is absorbed by antigen presenting cell (APC), generally present by MHC II, be difficult to produce cytotoxic lymphocyte (CTL), cannot eliminate the pathogen of invading cell; And part Ag is also lowered efficiency by non-APC picked-up.2) Ag is easy to inoculation position inactivation.3) mucus of continuous secretion not only hinders APC picked-up Ag but also flows, upgrades comparatively fast, often causes vaccine not enter i.e. a large amount of loss of body.
Summary of the invention
The object of the invention is to overcome the deficiency of existing vaccine, provide a kind of lipid A to modify the microneedle array vaccine adjuvant transmission system of vector construction.
The present invention is achieved by the following technical solutions:
Lipid A is modified a microneedle array vaccine adjuvant transmission system for vector construction, and described microneedle array comprises substrate and some micropins that is fixed on described substrate; Its composition of micropin described in each comprises lipid A modification carrier, auxiliary material and vaccine composition, forms vaccine adjuvant transmission system.
Microneedle array comprises substrate and some micropins that is fixed on substrate; Its composition of each micropin comprises lipid A modification carrier (comprising vaccine composition) and auxiliary material (excipient).
Lipid A is modified carrier, and lipid A comprises monophogphoryl lipid A, lipid A and lipopolysaccharide; Carrier comprises monolayer, few layer, multilamellar and multivesicular liposome, lipoid, microcapsule, or nanoparticle; Carrier body lotus positive electricity, negative electricity or neutrality.
As the further optimization of such scheme, the substrate cross section of described microneedle array is square (the general length of side is less than 3 centimetres) or circular (general diameter is less than 3 centimetres), is constituted by saccharide, polyvidone class (PVP), starch based, cellulose family or this type of Biodegradable material (excipient).
As the further optimization of such scheme, described in be fixed on the micropin of substrate, number is generally greater than 3; Be highly 50-500 μ m; Spacing is 100-400 μ m; For the cylinder that vertebral body or top are vertebral body, bottom surface diameter range is 50-500 μ m.
As the further optimization of such scheme, vaccine carrier and vaccine adjuvant that described vaccine composition is selected from subunit vaccine antigenic material, toxoid, deactivation or attenuated pathogens, comprises antigen; The described vaccine carrier that comprises antigen is liposome (comprising monolayer, multilamellar, multivesicular liposome), lipoid, nanoparticle or the microcapsule that carries antigen; Described vaccine adjuvant is lipid A, monophogphoryl lipid A, LPS (lipopolysaccharide), CpG-ODN, aluminum salt or pilin.
As the further optimization of such scheme, described vaccine composition or be encapsulated in described lipid A and modify in carrier or be adsorbed in described lipid A and modify carrier surface; Or described vaccine composition part is encapsulated in, and described lipid A is modified in carrier, part is adsorbed in described lipid A and modifies carrier or simply mix with carrier.
As the further optimization of such scheme, described lipid A modified liposome microneedle array vaccine adjuvant transmission system is inoculated or cutaneous inoculation by oral mucosa.
The preparation process of lipid A modified liposome microneedle array vaccine adjuvant transmission system mainly completes by poly-dimethoxy silane microneedle array mould.
(1) first prepare lipid A and modify carrier (or being loaded with vaccine composition);
(2) the lipid A modification carrier, the auxiliary element (or comprising vaccine composition) that are dispersed or dissolved in solvent are filled in the pin hole of microneedle array mould, then coated with substrate composition;
(3) the microneedle array mould after filling is placed under normal temperature condition to drying basin (containing desiccant such as anhydrous calcium chlorides, phosphorus pentoxide) is dry to be removed moisture or remove moisture by lyophilization, peel off, obtain described a kind of lipid A and modify carrier microneedle array vaccine.
Compared with prior art, the present invention is different from traditional mucosal vaccine, is also different from existing micropin vaccine (all by cutaneous inoculation), and the present invention overcomes the short length that has again the two concurrently of the two; The novel microneedle array vaccine system that is applicable to Different Kinds of Pathogens isoantigen, has the following advantages:
(1) applied widely, lipid A of the present invention is modified carrier microneedle array vaccine adjuvant transmission system can comprise the vaccine of different vaccine compositions formation for different pathogens; Vaccine composition can be antigen, attenuation or inactivating pathogens;
(2) stability is high, and it is solid preparation that lipid A of the present invention is modified carrier microneedle array vaccine adjuvant transmission system, and stability is high; MAV can protect antigen to exempt from (inoculation) surrounding material and destroy, external stability in reinforcement; Be expected to become de-cold chain or temperature control chain and be suitable for vaccine.
(3) safe, material therefor has good biocompatibility, and envelope antigen material obtains subunit vaccine, has higher-security.
(4) immune induction effect is strong, lipid A is modified carrier microneedle array vaccine by cutaneous inoculation, convenient and painless, can activate skin Langerhans cell (Langerhans cell), induction body produces antigen-specific immune response, the invasion of defence pathogen; Lipid A is modified carrier microneedle array vaccine and is inoculated by oral mucosa, can avoid vaccine to run off with saliva, also can eliminate oral mucosa multiple layer squamous cell and the surperficial mucus inhibition for vaccine picked-up, improve picked-up and the utilization ratio of antigen presenting cell for vaccine, effectively activate body immune system.
(5) set up multiple defense pathogen, lipid A is modified carrier microneedle array vaccine adjuvant transmission system and is inoculated and can be activated body immune system by oral mucosa, can either produce systemic immunity and reply, also can produce mucosal immune response, to pathogen, invasion forms dual defensive barrier.By cutaneous inoculation, convenient and painless, can activate skin Langerhans cell (Langerhans cell), induction body produces antigen-specific immune response, the invasion of defence pathogen.
In a word, lipid A of the present invention is modified microneedle array vaccine (the Microneedle Array Vaccine of vector construction, MAV), also be a kind of vaccine adjuvant transmission system (VacDAS), form safe, efficient, the stable vaccine for different pathogens by the different vaccine compositions of load.Microneedle array vaccine of the present invention is by skin or oral mucosa is convenient, painless, compliance is good.Especially by mucosal vaccination, microneedle array vaccine had both overcome the defect that existing micropin vaccine cannot mucosa immunity-inducing be replied, and had overcome again many deficiencies such as mucus obstruction, ingredients from lossing, antigen transmission efficiency that mucosal vaccination exists are low.Microneedle array vaccine of the present invention is by cutaneous inoculation, convenient and painless, can activate skin Langerhans cell, and induction body produces antigen-specific immune response, the invasion of defence pathogen.These two kinds of inoculation forms are all safer simultaneously.In addition,, because lipid A modification carrier micropin vaccine is anhydrous, stability is high, is expected to be applicable to temperature control chain, is conducive to generally inoculate.The present invention is that microneedle array vaccine is explored inoculation new way, for Oral inoculation is sought novel vaccine, also for Development of Novel CTC-VacDAS lays the foundation, has important scientific research meaning and clinical value widely.
Brief description of the drawings
Fig. 1 is the structural representation that lipid A of the present invention is modified the micropin of carrier (taking liposome as example) structure.
Detailed description of the invention
Below embodiments of the invention are elaborated, the present embodiment is implemented under taking technical solution of the present invention as prerequisite, has provided detailed description of the invention and operating process, but protection scope of the present invention is not limited to following embodiment.
Referring to Fig. 1, for lipid A of the present invention is modified the structural representation of the micropin that carrier (taking liposome as example) builds.First prepare lipid A and modify carrier as antigen vectors, then build microneedle array with described carrier and excipient, form vaccine adjuvant transmission system.
[embodiment 1]
< lipid A is modified microneedle array vaccine (MAV) > that multilamellar liposome builds
Taking OVA as antigen, with SPC/LA (100:1, mole ratio) be film material, total fat/OVA (20:1, mass ratio), as water, prepare liposome with film dispersion method taking 10% trehalose, 20%PVPk30 (excipient) solution, forming mean diameter is the OVA/Lipid A-liposome (OVA/LL) that 250 nanometers, zeta potential 6mV, envelop rate are 10%.Afterwards LL is mixed to (1:5 with aluminum phosphate (mean diameter 500 nanometers), W/W), be packed into (5 × 5 hole) pin hole of the microneedle array mould of being prepared by poly-dimethoxy silane by decompression, again coated with 10% trehalose, 20%PVPk30 (excipient) solution, lyophilizing dewaters again, strip off, obtain OVA/LL-MAV (6 × 6 micropins, substrate 0.65 × 0.65cm
2, be fixed on every micropin in substrate: 250 × 250 × 500 microns
3tetragonal pyramid needle body).After OVA/LL-MAV stores 2 weeks, after aquation, recover H1sAg-MLL, These parameters is without significant change.OVA/LL-MAV stored after 2 weeks,, compared with blank to mouse inoculation by oral mucosa, produced high-level OVA specific antibody and CTL after 3 weeks; Higher IgG1/IgG2a and high-level IFN-γ, show to inoculate Mus and produce Th1/Th2 mixed type immunne response; High-level IgA all detected at mice saliva, small intestinal flushing liquor, birth canal flushing liquor, show that mice had both produced systemic immunity and replied, also produced mucosal immune response simultaneously.(OVA, ovalbumin; LPS, lipopolysaccharide; SPC, fabaceous lecithin phatidylcholine; LA, lipid A, lipid A; MPC, mannose-PEG2000-cholesterol; DOTAP, 1,2-dioleoyl-3-trimethylammonium-propane, 1,2-dioleoyl-N, N, N-trimethyl-propylamine; PVPk30, polyvidone).
[embodiment 2]
The microneedle array vaccine > that the dual modified liposome of < mannose group/lipid A (lotus positive electricity) builds
With influenza A H1N1 influenza virus (influenza A (H1N1) virus) surface antigen (hemagglutinin1 antigen, H1sAg) be antigen, with SPC/MPC/MPLA/DOTAP (20:1:0.05:1, mole ratio) be film material, total fat/H1sAg (20:1, mass ratio), taking 10% trehalose, 20%PVPk30 (excipient) solution as water, prepare liposome with reverse evaporation, forming mean diameter is the dual modified liposome of MPC/Lipid A (MLL) that 320 nanometers, zeta potential 13mV, envelop rate are 59%.Afterwards MLL is packed into (6 × 6 hole) pin hole of the microneedle array mould of being prepared by poly-dimethoxy silane by decompression, again coated with 10% trehalose, 20%PVPk30 (excipient) solution, again mould is inserted to anhydrous calcium chloride drying basin dry 8 hours, strip off, obtain H1sAg-MLL-MAV (6 × 6 micropins, substrate 0.65 × 0.65cm
2, be fixed on every micropin in substrate: 100 × 100 × 3.14 × 500/3 micron
3cone micropin).After H1sAg-MLL-MAV stores 2 weeks, after aquation, recover H1sAg-MLL, These parameters is without significant change.H1sAg-MLL-MAV stored after 2 weeks,, compared with blank to mouse inoculation by oral mucosa, produced high-level H1sAg specific antibody and CTL after 3 weeks; Higher IgG1/IgG2a and high-level IFN-γ, show to inoculate Mus and produce Th1/Th2 mixed type immunne response; High-level IgA all detected at mice saliva, small intestinal flushing liquor, birth canal flushing liquor, show that mice had both produced systemic immunity and replied, also produced mucosal immune response simultaneously.(H1sAg, influenza A H1N1 influenza virus surface antigen; SPC, fabaceous lecithin phatidylcholine; MPLA, monophosphoryl lipid A, single phospholipid A; MPC, mannose-PEG2000-cholesterol; DOTAP, 1,2-dioleoyl-3-trimethylammonium-propane, 1,2-dioleoyl-N, N, N-trimethyl-propylamine; PVPk30, polyvidone).
[embodiment 3]
< lipid A is modified the microneedle array Hepatitis B virus vaccine > that multivesicular liposome builds
Taking HBsAg as antigen, with SPC/GMO/LPS (20:4:0.05, mole ratio) be film material, total fat/HBsAg (20:1, mass ratio), as water, prepare multivesicular liposome with emulsifying-evaporation taking 10% sucrose, 30%PVPk17 (excipient) solution, forming mean diameter is that the LPS that 520 nanometers, zeta potential-12mV, envelop rate are 72% modifies multivesicular liposome (LML).Afterwards MLL is packed into (6 × 6 hole) pin hole of the microneedle array mould of being prepared by poly-dimethoxy silane by decompression, again coated with 10% sucrose, 30%PVPk17 (excipient) solution, again mould is inserted to anhydrous calcium chloride drying basin dry 8 hours, strip off, obtain HBsAg-LML-MAV (8 × 8 micropins, substrate 0.75 × 0.75cm
2, be fixed on every micropin in substrate: 250 × 250 × 500 microns
3tetragonal prism needle body+250 × 250 × 50/3 micron
3tetragonal pyramid needle point).After HBsAg-LML-MAV stores 3 days, after aquation, recover HBsAg-LML, These parameters is without significant change.HBsAg-LML-MA, compares with blank to mouse inoculation by oral mucosa, produces high-level HBsAg specific antibody and CTL after 3 weeks; Higher IgG1/IgG2a and high-level IFN-γ, show to inoculate Mus and produce Th1/Th2 mixed type immunne response; High-level IgA all detected at mice saliva, small intestinal flushing liquor, birth canal flushing liquor, show that mice had both produced systemic immunity and replied, also produced mucosal immune response simultaneously.(HBsAg, hepatitis B virus surface antigen; LPS, lipopolysaccharide; SPC, fabaceous lecithin phatidylcholine; GL, glycerin, glycerol; GMO, glyceryl monooleate, glyceryl monooleate LA, lipid A, lipid A; MPC, mannose-PEG2000-cholesterol; DOTAP, 1,2-dioleoyl-3-trimethylammonium-propane, 1,2-dioleoyl-N, N, N-trimethyl-propylamine; PVPk30, polyvidone).
[embodiment 4]
The microneedle array vaccine > of <H1sAg-lipid A modified liposome
With SPC/MPLA/DOTAP (20:0.05:1, mole ratio) be film material, taking influenza A H1N1 influenza virus (influenza A (H1N1) virus) surface antigen (H1sAg), 10% trehalose, 20%PVPk30 (excipient) solution as water, total fat/flu virus (20:1, mass ratio), prepare liposome with reverse evaporation, the LL that form that mean diameter is 360, zeta potential 13mV, envelop rate is 53%).Afterwards flu-LL is packed into (9 × 9 hole) pin hole of the microneedle array mould of being prepared by poly-dimethoxy silane by decompression, again coated with 10% trehalose, 20%PVPk30 (excipient) solution, again mould is inserted to anhydrous calcium chloride drying basin dry 8 hours, strip off, obtain flu-LL-MAV (9 × 9 micropins, substrate 0.85 × 0.85cm
2, be fixed on every micropin in substrate: 100 × 100 × 3.14 × 500/3 micron
3cone micropin).After flu-LL-MAV stores 2 weeks, after aquation, recover flu-LL, These parameters is without significant change.Flu-LL-MAV stored after 2 weeks,, compared with blank to mouse inoculation by lagging skin, produced high-level H1sAg specific antibody and CTL after 3 weeks; Higher IgG1/IgG2a and high-level IFN-γ, show to inoculate Mus and produce Th1/Th2 mixed type immunne response.(H1sAg, influenza A H1N1 influenza virus surface antigen; SPC, fabaceous lecithin phatidylcholine; MPLA, monophosphoryl lipid A, single phospholipid A; MPC, mannose-PEG2000-cholesterol; DOTAP, 1,2-dioleoyl-3-trimethylammonium-propane, 1,2-dioleoyl-N, N, N-trimethyl-propylamine; PVPk30, polyvidone).
[embodiment 5]
< lipid A modify microcapsule build microneedle array vaccine >
Taking H1sAg as antigenic component, taking gelatin/arabic gum/LPS as capsule material, adopt complex coacervation to prepare microcapsule, be scattered in 10% trehalose, 20%PVPk30 (excipient) solution.Be packed into afterwards (6 × 6 hole) pin hole of the microneedle array mould of being prepared by poly-dimethoxy silane by decompression, then coated with 10% sucrose, 20%PVPk30 solution, then mould is inserted to anhydrous calcium chloride drying basin dry 10 hours; Or remove moisture by lyophilizing, and strip off, obtain OVA-MAV (6 × 6 micropins, substrate 0.65 × 0.65cm
2, be fixed on every micropin in substrate: 250 × 250 × 500 microns
3needle body+250 × 250 × 50 micron
3needle point).Give mouse inoculation by oral mucosa subsequently, microexamination shows, micropin needle body can sting and penetrate mucosa.Compare with blank, after surrounding, mice produces high-level H1Ag specific antibody and CTL; High-level IgA all detected at mice saliva, small intestinal flushing liquor, birth canal flushing liquor, show that mice had both produced systemic immunity and replied, also produced mucosal immune response simultaneously.The mice of inoculation H1Ag-MLL gives lethal dose influenza A H1N1 influenza virus, and 100% survives after 1 week, and Mice Inoculated does not give lethal dose influenza A H1N1 influenza virus and only survives 10%.
[embodiment 6]
The microneedle array vaccine > that < lipoid adjuvant builds
The MAV that lipoid (niosome) adjuvant-transmission system builds: taking OVA as antigen, MPLA, Span60/LA (20:1:0.05:1, mole ratio) be film material, total fat/H1sAg (20:1, mass ratio), taking 10% trehalose, 20%PVPk30 (excipient) solution as water, prepare lipoid vesicle with reverse evaporation, forming mean diameter is the lipoid that 300 nanometers, zeta potential-25mV, envelop rate are 63%.Afterwards lipoid is packed into (8 × 8 hole) pin hole of the microneedle array mould of being prepared by poly-dimethoxy silane by decompression, insert anhydrous calcium chloride drying basin dry 8 hours again coated with 10% sucrose, 20%PVPk30 (excipient) solution, then by microneedle array device; Or dewater by freeze-dried, strip off, obtain H1sAg-niosome-MAV (8 × 8 micropins, substrate 0.75 × 0.75cm
2, be fixed on every micropin in substrate: 250 × 250 × 500 microns
3needle body+250 × 250 × 50 micron
3needle point).After H1sAg-niosome-MAV stores 2 weeks, after aquation, recover H1sAg-niosomes, These parameters is without significant change.H1sAg-niosome-MAV stored after 2 weeks, and by oral mucosa, to mouse inoculation, microexamination shows, micropin needle body can sting and penetrate mucosa.Compare with blank, after surrounding, mice produces high-level OVA specific antibody and CTL; Higher IgG1/IgG2a and high-level IFN-γ, show to inoculate Mus and produce Th1/Th2 mixed type immunne response; High-level OVA specificity IgA all detected at mice saliva, small intestinal flushing liquor, birth canal flushing liquor, show that mice had both produced systemic immunity and replied, also produced mucosal immune response simultaneously.(Span60, sorbester p18; LA, lipid A, lipid A).
[embodiment 7]
< solid lipid nanoparticle adjuvant build microneedle array vaccine >
Taking OVA as antigen, taking lipid A as adjuvant, taking SPC/GMS/LA as material, adopt emulsifying-rotary evaporation legal system for solid lipid nanoparticle, total fat/H1sAg (20:1, mass ratio), be scattered in 10% sucrose, 20%PVPk30 (excipient) solution, forming mean diameter is the solid lipid nanoparticle (SLNs) that 200 nanometers, zeta potential-31mV, envelop rate are 42%.Afterwards SLNs is packed into (10 × 10 hole) pin hole of the microneedle array mould of being prepared by poly-dimethoxy silane by decompression, insert anhydrous calcium chloride drying basin dry 8 hours again coated with 10% sucrose, 20%PVPk30 (excipient) solution, then by microneedle array mould; Or dewater by freeze-dried, strip off, obtain H1sAg-SLN-MAV (10 × 10 micropins, substrate 0.85 × 0.85cm
2, be fixed on every micropin in substrate: 250 × 250 × 500 microns
3needle body+250 × 250 × 50 micron
3needle point).After H1sAg-SLN-MAV stores 2 weeks, after aquation, recover H1sAg-niosomes, These parameters is without significant change.H1sAg-SLN-MAV stored after 2 weeks, shaved mao rear skin to mouse inoculation by back, and microexamination shows, micropin needle body can sting and penetrate mouse skin epidermis.Compare with blank, after surrounding, mice is compared with blank, produces high-level OVA specific antibody and CTL after 3 weeks; Higher IgG1/IgG2a and high-level IFN-γ, show to inoculate Mus and produce Th1 type immunne response.(GMS, glyceryl monostearate).
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.
Claims (10)
1. lipid A is modified the microneedle array vaccine adjuvant transmission system of vector construction, it is characterized in that: microneedle array comprises substrate and some micropins that is fixed on substrate; Described in each, its composition of micropin comprises lipid A modification carrier, auxiliary material and vaccine composition, forms vaccine adjuvant transmission system.
2. lipid A according to claim 1 is modified the microneedle array vaccine adjuvant transmission system of vector construction, it is characterized in that: described substrate is constituted by saccharide, polyvidone class (PVP), starch based, cellulose family or these Biodegradable materials.
3. lipid A according to claim 1 is modified the microneedle array vaccine adjuvant transmission system of vector construction, it is characterized in that: described some micropins that are fixed on substrate, and number is generally greater than 3; Be highly 50-500 μ m; Spacing is 100-400 μ m; For the cylinder that vertebral body or top are vertebral body, diameter range is 50-500 μ m.
4. lipid A according to claim 1 is modified the microneedle array vaccine adjuvant transmission system of vector construction, it is characterized in that: described lipid A is modified carrier, and lipid A comprises monophogphoryl lipid A, lipid A and lipopolysaccharide; Described carrier comprises monolayer, few layer, multilamellar or multivesicular liposome, lipoid, microcapsule, or nanoparticle; Carrier lotus positive electricity, negative electricity or neutrality.
5. lipid A according to claim 1 is modified the microneedle array vaccine adjuvant transmission system of vector construction, it is characterized in that: described lipid A is modified carrier, can further adopt functional molecular to modify, for example, modify carrier with mannose group, improve antigen transmission object to reach.
6. lipid A according to claim 1 is modified the microneedle array vaccine adjuvant transmission system of vector construction, it is characterized in that: described auxiliary material be can give described micropin intensity, hardness, shape excipient, select saccharide (such as sucrose, trehalose, lactose etc.), PVP class, starch based, cellulose family, or the combination of these materials; Described auxiliary material also comprise other vaccine adjuvants (such as aluminum salt, CpG-ODN, CpG-ODN, saponin, Squalene, imiquimod etc. and pilin etc.), to improve vaccine adjuvant function.
7. lipid A according to claim 1 is modified the microneedle array vaccine adjuvant transmission system of vector construction, it is characterized in that: described vaccine composition is selected from subunit vaccine antigenic material, toxoid, deactivation or attenuated pathogens.
8. lipid A according to claim 1 is modified the microneedle array vaccine adjuvant transmission system of vector construction, it is characterized in that: described vaccine composition or be encapsulated in described lipid A and modify in carrier, or be adsorbed in described lipid A modification carrier surface, or be and the simple mechanical mixture of carrier.
9. lipid A according to claim 1 is modified the microneedle array vaccine adjuvant transmission system of vector construction, it is characterized in that: described lipid A modified liposome microneedle array vaccine adjuvant transmission system is mainly inoculated or cutaneous inoculation by oral mucosa.
10. lipid A claimed in claim 1 is modified the microneedle array vaccine adjuvant transmission system of vector construction, it is characterized in that, its preparation method mainly relies on and adopts microneedle array reverse tool prepared by poly-dimethoxy silane to realize, and comprises the steps: that (1) first prepare lipid A and modify carrier (or being loaded with vaccine composition);
(2) lipid A that is dispersed or dissolved in solvent (aqueous solution) is modified to carrier, auxiliary element and vaccine composition and be filled in the pin hole of microneedle array mould, then coated with substrate composition (aqueous solutions of above-mentioned auxiliary material);
(3) the microneedle array mould after filling is placed under normal temperature condition to drying basin (containing desiccant such as anhydrous calcium chlorides, phosphorus pentoxide) is dry to be removed moisture or remove moisture by lyophilization, strip off, obtain described lipid A and modify microneedle array vaccine.
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| CN107206065A (en) * | 2014-12-22 | 2017-09-26 | 博莱科瑞士股份有限公司 | The micro-capsule of gas filling as vaccine |
| KR20180123695A (en) * | 2016-04-15 | 2018-11-19 | 후지필름 가부시키가이샤 | Micro needle array |
| CN108969771A (en) * | 2018-08-07 | 2018-12-11 | 中国医学科学院生物医学工程研究所 | The total load antigen of mannose-modified and double immune agonist phosphatide hybridized polymer vesicas and the preparation method and application thereof |
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| CN104367567A (en) * | 2014-10-22 | 2015-02-25 | 北京化工大学 | Extrusion embossing based quick vaccine patch and preparation method thereof |
| CN107206065A (en) * | 2014-12-22 | 2017-09-26 | 博莱科瑞士股份有限公司 | The micro-capsule of gas filling as vaccine |
| CN109069817B (en) * | 2016-04-15 | 2021-07-13 | 富士胶片株式会社 | Microneedle array |
| KR20180123695A (en) * | 2016-04-15 | 2018-11-19 | 후지필름 가부시키가이샤 | Micro needle array |
| CN109069817A (en) * | 2016-04-15 | 2018-12-21 | 富士胶片株式会社 | Microneedle array |
| JPWO2017179614A1 (en) * | 2016-04-15 | 2019-02-21 | 富士フイルム株式会社 | Microneedle array |
| EP3444004A4 (en) * | 2016-04-15 | 2019-03-27 | Fujifilm Corporation | Microneedle array |
| AU2017251040B2 (en) * | 2016-04-15 | 2019-07-18 | Fujifilm Corporation | Microneedle array |
| KR102194109B1 (en) * | 2016-04-15 | 2020-12-22 | 후지필름 가부시키가이샤 | Microneedle array |
| US11266822B2 (en) | 2016-04-15 | 2022-03-08 | Fujifilm Corporation | Microneedle array |
| CN108969771A (en) * | 2018-08-07 | 2018-12-11 | 中国医学科学院生物医学工程研究所 | The total load antigen of mannose-modified and double immune agonist phosphatide hybridized polymer vesicas and the preparation method and application thereof |
| CN108969771B (en) * | 2018-08-07 | 2021-06-29 | 中国医学科学院生物医学工程研究所 | Mannose-modified co-carried antigen and double immune agonist phospholipid hybrid polymer vesicle and preparation method and application thereof |
| CN112516452A (en) * | 2020-12-18 | 2021-03-19 | 南京鼓楼医院 | Frozen microneedle array and preparation method and application thereof |
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